Scaffold-free self-organized 3D synthetic tissue

• US 9,694,107 B2
• Filed: 05/26/2016
• Issued: 07/04/2017
• Est. Priority Date: 08/01/2003
• Status: Active Grant

• Current Assignee: Two Cells Co. Ltd.
• Original Assignee: Two Cells Co. Ltd.
• Inventors: Nakamura, Norimasa, Yoshikawa, Hideki, Ando, Wataru
• Primary Examiner(s): DAVIS, RUTH A
• Application Number:
  US15/166,059
• Publication Number:
  US 20160256608A1
• Time in Patent Office: 404 Days
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61L 2430/06 for cartilage reconstruction, e.g. meniscus
• CPC Class Codes: A61L 27/24 Collagen
• CPC Class Codes: A61L 27/3612 Cartilage, synovial fluid
• CPC Class Codes: A61L 27/3633 Extracellular matrix [ECM]
• CPC Class Codes: A61L 27/3817 Cartilage-forming cells, e.g. pre-chondrocytes
• CPC Class Codes: A61L 27/3852 Cartilage, e.g. meniscus
• CPC Class Codes: A61L 27/3895 using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
• CPC Class Codes: A61L 27/50 Materials characterised by their function or physical properties , e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2501/135 Platelet-derived growth factor [PDGF]
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2501/20 Cytokines; Chemokines
• CPC Class Codes: C12N 2501/33 Insulin together with transferrin C12N2500/25; Insulin-like growth factors C12N2501/105
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 2501/999 : Small molecules not provided for elsewhere
• CPC Class Codes: C12N 2502/1311 : Osteocytes, osteoblasts, odontoblasts
• CPC Class Codes: C12N 2506/1384 : from adipose-derived stem cells [ADSC], from adipose stromal stem cells
• CPC Class Codes: C12N 5/0654 : Osteocytes, Osteoblasts, Odontocytes; Bones, Teeth
• CPC Class Codes: C12N 5/0655 : Chondrocytes; Cartilage

Abstract

The present invention can be used for actual implantation surgery without a scaffold. The present invention provides a synthetic tissue or complex which can be produced by culture and has a high level of differentiation ability. The present invention also provides a therapy and medicament for repairing and/or regenerating tissue using replacement and covering. By culturing cells under specific culture conditions such that medium contains an extracellular matrix synthesis promoting agent, the cells are organized and are easily detached from a culture dish. The present invention was achieved by finding such a phenomenon. In addition, the self contraction of the tissue can be regulated by culturing the tissue in a suspended manner. Therefore, it is possible to regulate the three-dimensional shape of the tissue. The present invention also provides a method for producing an implantable synthetic tissue which does not require a plurality of monolayer cell sheets assembled to form a three-dimensionally structured synthetic tissue. The present invention is characterized by richness in adhesion molecules, nonnecessity of additional fixation at an implantation site, and good biological integration.

Claims

1. A tissue complex comprising an implantable synthetic autograft or allograft tissue and another synthetic tissue which is different from the implantable autograft or allograft tissue,

wherein the implantable synthetic autograft or allograft tissue is substantially made of (i) cells that are autologous or allogeneic relative to the recipient and which thereby constitute the synthetic autograft or allograft tissue, said cells being selected from the group consisting of myoblasts, mesenchymal stem cells, adipocytes, synovial cells and bone marrow cells;

and (ii) an extracellular matrix (ECM) derived from the cells constituting the synthetic autograft or allograft tissue, said extracellular matrix comprising fibronectin, collagen I, collagen III, and vitronectin,

wherein the synthetic autograft or allograft tissue is free of scaffolds and comprises multiple layers of said cells, and

wherein the implantable synthetic autograft or allograft tissue has integration ability with its surroundings when implanted and has sufficient strength to provide self-supporting ability.

2. The tissue complex of claim 1, wherein the fibronectin, collagen I, collagen III, and vitronectin are evenly distributed in the synthetic autograft or allograft tissue.

3. The tissue complex of claim 1, wherein the fibronectin, collagen I, collagen III, and vitronectin three-dimensionally wrap the cells.

4. The tissue complex of claim 1, wherein the extracellular matrix is integrated throughout the tissue.

5. The tissue complex of claim 1, wherein the implantable synthetic autograft or allograft tissue comprises 10 or more layers of said cells.

6. The tissue complex of claim 1, wherein the implantable synthetic autograft or allograft tissue has a thickness of at least 50 μ

m and is dimensioned so as to fit an injured site in a tissue or organ.

7. The tissue complex of claim 1, wherein the another synthetic tissue is an artificial bone.

8. The tissue complex of claim 7, wherein the artificial bone is made of hydroxyapatite.

9. The tissue complex of claim 1, wherein the another synthetic tissue is a microfibrous collagen medical device, a collagen gel, a hyaluronic acid gel, an agarose gel, an alginate gel or beads.